PMID- 38189517
OWN - NLM
STAT- MEDLINE
DCOM- 20240109
LR  - 20240109
IS  - 1940-087X (Electronic)
IS  - 1940-087X (Linking)
IP  - 202
DP  - 2023 Dec 22
TI  - Immunomagnetic Isolation of the Vascular Wall-Resident CD34+ Stem Cells from 
      Mice.
LID - 10.3791/66193 [doi]
AB  - Resident CD34(+) vascular wall-resident stem and progenitor cells (VW-SCs) are 
      increasingly recognized for their crucial role in regulating vascular injury and 
      repair. Establishing a stable and efficient method to culture functional murine 
      CD34(+) VW-SCs is essential for further investigating the mechanisms involved in 
      the proliferation, migration, and differentiation of these cells under various 
      physiological and pathological conditions. The described method combines magnetic 
      bead screening and flow cytometry to purify primary cultured resident CD34(+) 
      VW-SCs. The purified cells are then functionally identified through 
      immunofluorescence staining and Ca(2+) imaging. Briefly, vascular cells from the 
      adventitia of the murine aorta and mesenteric artery are obtained through tissue 
      block attachment, followed by subculturing until reaching a cell count of at 
      least 1 x 10(7). Subsequently, CD34(+) VW-SCs are purified using magnetic bead 
      sorting and flow cytometry. Identification of CD34(+) VW-SCs involves cellular 
      immunofluorescence staining, while functional multipotency is determined by 
      exposing cells to a specific culture medium for oriented differentiation. 
      Moreover, functional internal Ca(2+) release and external Ca(2+) entry is 
      assessed using a commercially available imaging workstation in Fura-2/AM-loaded 
      cells exposed to ATP, caffeine, or thapsigargin (TG). This method offers a stable 
      and efficient technique for isolating, culturing, and identifying vascular 
      wall-resident CD34(+) stem cells, providing an opportunity for in vitro studies 
      on the regulatory mechanisms of VW-SCs and the screening of targeted drugs.
FAU - Han, Chuting
AU  - Han C
AD  - Key Laboratory of Medical Electrophysiology of Ministry of Education and Medical 
      Electrophysiological Key Laboratory of Sichuan Province, Institute of 
      Cardiovascular Research, Southwest Medical University.
FAU - Xie, Cheng
AU  - Xie C
AD  - Key Laboratory of Medical Electrophysiology of Ministry of Education and Medical 
      Electrophysiological Key Laboratory of Sichuan Province, Institute of 
      Cardiovascular Research, Southwest Medical University.
FAU - Dang, Qingya
AU  - Dang Q
AD  - Key Laboratory of Medical Electrophysiology of Ministry of Education and Medical 
      Electrophysiological Key Laboratory of Sichuan Province, Institute of 
      Cardiovascular Research, Southwest Medical University.
FAU - Zhang, Xiaolin
AU  - Zhang X
AD  - Key Laboratory of Medical Electrophysiology of Ministry of Education and Medical 
      Electrophysiological Key Laboratory of Sichuan Province, Institute of 
      Cardiovascular Research, Southwest Medical University.
FAU - Li, Chunshu
AU  - Li C
AD  - Key Laboratory of Medical Electrophysiology of Ministry of Education and Medical 
      Electrophysiological Key Laboratory of Sichuan Province, Institute of 
      Cardiovascular Research, Southwest Medical University.
FAU - Yang, Yan
AU  - Yang Y
AD  - Key Laboratory of Medical Electrophysiology of Ministry of Education and Medical 
      Electrophysiological Key Laboratory of Sichuan Province, Institute of 
      Cardiovascular Research, Southwest Medical University.
FAU - Cheng, Jun
AU  - Cheng J
AD  - Key Laboratory of Medical Electrophysiology of Ministry of Education and Medical 
      Electrophysiological Key Laboratory of Sichuan Province, Institute of 
      Cardiovascular Research, Southwest Medical University; lzcj1221@swmu.edu.cn.
FAU - Li, Pengyun
AU  - Li P
AD  - Key Laboratory of Medical Electrophysiology of Ministry of Education and Medical 
      Electrophysiological Key Laboratory of Sichuan Province, Institute of 
      Cardiovascular Research, Southwest Medical University; lpyun@swmu.edu.cn.
LA  - eng
PT  - Journal Article
PT  - Video-Audio Media
DEP - 20231222
PL  - United States
TA  - J Vis Exp
JT  - Journal of visualized experiments : JoVE
JID - 101313252
RN  - 0 (Antigens, CD34)
RN  - 0 (Cell Adhesion Molecules)
SB  - IM
MH  - Animals
MH  - Mice
MH  - *Stem Cells
MH  - Adventitia
MH  - *Vascular System Injuries
MH  - Aorta
MH  - Antigens, CD34
MH  - Cell Adhesion Molecules
EDAT- 2024/01/08 12:42
MHDA- 2024/01/09 06:42
CRDT- 2024/01/08 09:53
PHST- 2024/01/09 06:42 [medline]
PHST- 2024/01/08 12:42 [pubmed]
PHST- 2024/01/08 09:53 [entrez]
AID - 10.3791/66193 [doi]
PST - epublish
SO  - J Vis Exp. 2023 Dec 22;(202). doi: 10.3791/66193.